LOCUS       sequence-307309-        7305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 M (membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7305)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7305)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133569
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    45..116
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(121..167)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    185..233
                     /note="synthetic polyadenylation signal"
     enhancer        314..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      890..899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1562..1582
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1583..1660
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1729..1850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1740..1759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1794..1813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1965..2011
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2041..2177
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2222..2293
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2519..3107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2599..2618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3607..4816
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4653..4672
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4820..5848
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5875..5923
                     /note="synthetic polyadenylation signal"
     CDS             join(6189..6257,6258..7049)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6407..6426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7062..7198
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
       61 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
      121 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
      181 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
      241 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      841 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg ccaccatggc
      901 agattccaac ggtactatta ccgttgaaga gcttaaaaag ctccttgaac aatggaacct
      961 agtaataggt ttcctattcc ttacatggat ttgtcttcta caatttgcct atgccaacag
     1021 gaataggttt ttgtatataa ttaagttaat tttcctctgg ctgttatggc cagtaacttt
     1081 agcttgtttt gtgcttgctg ctgtttacag aataaattgg atcaccggtg gaattgctat
     1141 cgcaatggct tgtcttgtag gcttgatgtg gctcagctac ttcattgctt ctttcagact
     1201 gtttgcgcgt acgcgttcca tgtggtcatt caatccagaa actaacattc ttctcaacgt
     1261 gccactccat ggcactattc tgaccagacc gcttctagaa agtgaactcg taatcggagc
     1321 tgtgatcctt cgtggacatc ttcgtattgc tggacaccat ctaggacgct gtgacatcaa
     1381 ggacctgcct aaagaaatca ctgttgctac atcacgaacg ctttcttatt acaaattggg
     1441 agcttcgcag cgtgtagcag gtgactcagg ttttgctgca tacagtcgct acaggattgg
     1501 caactataaa ttaaacacag accattccag tagcagtgac aatattgctt tgcttgtaca
     1561 ggaaaacctt tactttcagt cgaagaggcg atggaaaaag aattttatcg cggtgagtgc
     1621 agccaacaga ttcaaaaaga tatcctcgtc tggtgccctc taaggatccg caggcctctg
     1681 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1741 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1801 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1861 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1921 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     1981 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     2041 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     2101 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     2161 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     2221 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     2281 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     2341 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     2401 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     2461 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2701 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2941 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3121 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3181 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3241 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3301 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     3361 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     3421 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3481 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3541 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3601 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3661 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3721 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     3781 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     3841 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     3901 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     3961 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     4021 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     4081 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     4141 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     4201 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     4261 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     4321 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     4381 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     4441 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     4501 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4561 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4621 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4681 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4741 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     4801 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     4861 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     4921 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     4981 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     5041 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     5101 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     5161 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     5221 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     5281 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     5341 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     5401 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     5461 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     5521 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     5581 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     5641 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     5701 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     5761 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     5821 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     5881 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     5941 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     6001 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     6061 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     6121 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     6181 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6241 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6301 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6361 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6421 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6481 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6541 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6601 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6661 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6721 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     6781 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     6841 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6901 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6961 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7021 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     7081 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     7141 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     7201 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     7261 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtc
//
